Chapter 9 Two Simple Electrophysiological Preparations Using Grasshoppers Charles Carlson, Bonnie Jones, Wayne La Rochelle with Illustrations and Photographs by Susan Schwartzenberg Exploratorium 3601 Lyon Street San Francisco, California 94123 Charles Carlson is director of the Life Sciences section of the Ex- ploratorium, a science museum in San Francisco. Born and raised in the San Francisco Bay Area, he attended the University of Cal- ifornia at Berkeley, from which he graduated with a B.S. in zoology and an A.B. in communications and public policy in 1970. Since coming to the Exploratorium in 1972, he has built more than 20 biology exhibits which are currently in use in the museum. Bonnie M. Jones graduated with honors from the University of California, Davis (1974) with a B.S. in zoology. After an extensive trip through Europe, she attended the University of Michigan in a Ph.D. program. She returned to the San Francisco area and worked at the Exploratorium as a staff Biologist. In 1980 she grad- uated with a Master’s degree in environmental engineering from the University of California, Berkeley. During the last year of school, Bonnie Jones was employed half-time as a Research As- sistant on the Lawrence Berkeley Laboratory (LBL) Oil Shale Pro- ject, wastewater treatment program. Since graduation, she has been employed full-time as a LBL staff scientist on the Oil Shale Project. Ms. Jones resides in Marin county and maintains interests in hiking, bird watching and sewing. Wayne La Rochelle has been a staff biologist at the Exploratorium since 1976. He received a B.S. in zoology, with an emphasis on neurophysiology, from University of California at Davis in 1972. In 1976 he began his association with the Exploratorium by par- ticipating in a program to build exhibits demonstrating basic neu- robiological concepts to the public. 181 181 182 Electrophysiology of the Grasshopper Susan Schwartzenberg studied photography at the School of the Art Institute of Chicago and received in 1973 a Bachelor of Fine Arts degree and Bachelor of Science in Art Education from North- ern Illinois University. Since 1975 she has been illustrator and photographer for the Life Science section of the Exploratorium in San Francisco. I. Introduction Two long-lasting electrophysiological preparations utilizing grasshoppers are currently on display as live exhibits at the Exploratorium, a science mu- seum in San Francisco. The techniques for preparing these demonstrations, as well as the maintenance of the two grasshopper species, Schistocerca nitens and S. americana, are described in the hopes that they will be of use in the college biology laboratory. These preparations were not developed with lab- oratory teaching in mind, but it is hoped that they may be successfully mod- ified for use in that area. “The Watchful Grasshopper” The “Watchful Grasshopper” exhibit uses chronically implanted wire electrodes to record from the ventral nerve cord of a grasshopper. The neural activity which is detected by the electrodes and displayed via recording am- plifiers and oscilloscope clearly shows the electrical activity of two paired neurons; the Descending Contra-lateral Motion Detector (DCMD) and the Descending Ipsi-lateral Motion Detector (DIMD). These cells usually gen- erate nerve impulses in response to novel moving objects in the visual field and may show some activity in response to sounds. At the exhibit, visitors may explore and map the visual field of the grasshopper, determine what type of stimulus triggers impulses, and observe habituation to repeated stimulation in specific areas of the visual field. The preparation is of interest because motion detector cells are found in the visual systems of many other animals, including most vertebrates. They frequently serve to alert the animal to possible prey or danger. The motion detection system of the grasshopper is one of the most completely understood of these systems. “The Grasshopper Leg Twitch” The “Grasshopper Leg Twitch” exhibit uses the isolated leg of a grass- hopper to demonstrate that electrical stimulation can cause muscle contrac- tion. In this exhibit, visitors may vary the voltage to determine that there is a “threshold voltage” which must be attained before the leg muscles will contract. They may also change the frequency of electrical pulses administered to the leg to show that the shock-stimulus is cumulative. The teaching value Electrophysiology of the Grasshopper 183 of this preparation is limited by its use as a museum exhibit, whereas in a laboratory situation, there exists a much greater range for experimentation. For instance, measurements might be made of the strength of muscle con- traction, or the biomechanics of the knee joint and muscle attachment can be studied. Also, muscle tetanus might be studied, the position of the stimulating electrode could be changed, or the voltage applied to the leg could be modified much more drastically than is possible in the museum demonstration. This preparation lasts up to 24 hours with minimal care and may be used to show most of the phenomena demonstrated in other muscle-nerve preparations. Selection of Species Schistocerca nitens and S. americana were selected because of their rel- atively large body size. Also these animals are conveniently available to us at the Exploratorium from research laboratories at Stanford University and the University of California at Berkeley. With the animals we have received from these institutions, we have established and maintained grasshopper colonies with reasonable success. Most of the research on the grasshopper’s visual system has been con- ducted on Schistocerca nitens but our studies indicate that the DCMD and DIMD cells of this species are identical to those of S. americana. Since it is difficult to both ship and maintain either of these species, we recommend that locally available hoppers be used in these preparations whenever possible. Use of locally available species may require some field work, cage-construction, and exploration of your hopper’s neuroanatomy, but it does away with the bothersome need for numerous permits and the fabrication of exotic environ- ments. For example, in experimentation on species of grasshoppers indigenous to the San Francisco Bay Area, motion detector cells very similar to those found in S. nitens have been observed, Therefore, one should consider the information presented here only as a model around which a technique for preparing and maintaining other species of grasshoppers might be developed. II. The Solitary Desert Locust Natural History The genus Schistocerca contains solitary and gregarious species in both the Old and New Worlds. The species S. gregaria, which lives in northern and eastern Africa and western Asia, is the most destructive species of locust in the world. From the descriptions available, we know that the locust has plagued and devoured people’s food supplies since ancient times. Schistocerca nitens, the solitary desert locust with which we are most familiar in the United 184 Electrophysiology of the Grasshopper States, inhabits the arid regions of the southwestern United States and north- ern Mexico. These animals are considered an agricultural pest but do not swarm and are not as destructive as their larger African and Asian relatives. See Figure 9.1 Adult Schistocerca nitens have an extreme sexual size dimorphism. Gen- erally, females are about 6 cm from head to wing tip while the males are much smaller, being about 4 cm long. This difference is associated with the extra molt which females undergo. In the wild, the desert locust usually has one generation a year. Occa- sionally, during an extremely mild winter, a second generation may be pro- duced. The eggs are laid in loosely packed egg cases in the fall and stay in diapause through the winter. The young hatch with the spring greenery. The tiny (5 mm) hoppers immediately locate fleshy green plants and begin to feed. They undergo seven to eight molts, changing from a bright green wingless hopper to the intermediate unstriped beige morph, and finally to the winged brown adult with the characteristic leg striping and dark body pigmentation. Adults are capable of sustained flight, and it is only during the adult phase that mating and egg-laying occur. 1 eye grasshoppers have a keen sense of vision. They can almost see 360°. 2 antenna sensitive to touch and taste. You may see the insects cleaning their antennas to ensure good reception. 3 wings are used far escape and migration. 4 hind legs the powerful muscles enable the animal to jump long distances. 5 tympanic membrane sounds are sensed by movements of the membrane. 6 receptor hairs sensitive to touch and body position. Figure 9.1. Gross external anatomy of the grasshopper. Female-Schistocerca americana. Electrophysiology of the Grasshopper 185 The changing coloration of the growing locust provides an interesting study in adaptive coloration. All of the hoppers are green when they hatch and all of the adults the characteristic mottled brown or gray coloration. However, the intermediate instars are more variable. If the growing hoppers are crowded and reared in low humidity, they achieve dark coloration earlier in their lives than those raised in isolation and high humidity. It is thought that this coloration aids in camouflaging the animal from its natural predators. Dry, crowded conditions seem likely to dictate brown vegetation or exposed earth, either of which would favor a brown grasshopper. Conversely, humid, uncrowded conditions might favor a grasshopper which retains its green co- loration. In development, students should note the contrast between Schistocerca nitens and many familiar insects. The young hoppers are similar to the adults. They lack wings and genitalia and are a different color, but look and behave like the adults. This contrasts with the larval-pupal-adult life-cycle of the butterfly or moth. This difference in life-cycle strategies may be associated with the eating habits, food availability, and most propitious time of the plant- growing cycle for the larva or adult to exist.